Fig 1.

Localization of CAR^Ex8 is distinct from CAR^Ex7 and MAGI-1 localization in polarized epithelial cells, and CAR^Ex8 expression increases apical adenovirus infection. (A) Immunocytochemistry for endogenous MAGI-1 (green) and total CAR (red, Ab 1605p) in polarized MDCK cells. Overlapping staining was present at cell junctions (yellow, MERGE). (B) Endogenous CAR^Ex8 expression (red, Ab 5678p) is low in MDCK cells and does not overlap MAGI-1 (green). (C and D) Peptide competition for anti-CAR^Ex8 binding (red, Ab 5678p) demonstrates specificity of immunocytochemistry for CAR^Ex8 (C) but not total CAR (D) (red, Ab 1605p). (E) Immunocytochemistry for exogenous expression of GFP (green), CAR^Ex8 (red), or CAR^Ex7 (red) in polarized MDCK cells. Nuclei are stained with DAPI (blue). (F) Time course of transepithelial conductance in MDCK cells expressing exogenous GFP, CAR^Ex8, or CAR^Ex7 (representative). (G) Apical AdV-β-Gal transduction of 4-day-old polarized MDCK cells expressing exogenous GFP, CAR^Ex8, or CAR^Ex7 (data represent averages of the results of 3 separate experiments; n = 4 to 6 replicates per experiment). Representative single optical x-y or x-z plane confocal images are shown (60× oil immersion); dashed lines in panel E represent Millicell membrane. Bars = 20 μm. *, P < 0.05.